重组弓形虫14-3-3抗原用于弓形虫血清抗体的检测

Signaling protein of 14-3-3 as a diagnostic antigen for the detection of antibodies against Toxoplasma gondii

目的 探讨重组弓形虫信号转导蛋白14- 3- 3(Toxo14- 3- 3)在弓形虫病免疫诊断中的价值。方法 根据弓形虫Beverly株猫肠上皮细胞增殖阶段的14- 3 -3编码基因,设计合成引物,自RH株速殖子基因组克隆Toxo14- 3 -3的ORF,克隆pET 28a质粒,转化大肠杆菌,获得表达产物并纯化融合蛋白,以此抗原包被反应板进行ELISA检测,并与粗制抗原及市售试剂进行比较。结果 对400份血清标本的平行检测,Toxo14 -3 -3抗原、粗制抗原和市售试剂盒的阳性率分别为2 5%、3 0%和3 .25%,三者之间差异无显著性(P>0 .05);三者之间的符合率为99 0%、98.8%和99 .0%,差异无显著性(P>0. 05 )。结论 重组Toxo14 -3- 3抗原检测弓形虫血清抗体具有潜在价值。

Objective To investigate the potential value of Toxoplasma signaling protein of 14-3-3(Toxo14-3-3) in the diagnosis of toxoplasmosis. Methods Pimers were designed based on the open reading frame of Toxo14-3-3 in feline enteroepithelial stage of Toxoplasma. DNA fragments were cloned and ligated to expression vector of pET-28 a, followed by transformation of E.coli BL21.The fusion expression was purified by SDS-PAGE and used as diagnostic antigen in ELISA for the detection of antibodies against Toxoplasma in 400 outpatients. The results were compared in parallel with crude antigen(CA) prepared from RH strain tachyzoites and diagnostic agent kit commercially available. Results The positive rates of Toxoplasma antibodies tested by Toxo14-3-3,CA,and the kit were 2.5%,3.0%, and 3.25%,respectively.High accordance(99.0%,98.8%,and 99%) was found among the antigens used above. No significant difference was noted ( P >0.05). Conclusion The recombinant Toxo-14-3-3 has a potential value in the serodiagnosis of toxoplasmosis.